Perforation of metastatic melanoma to the small bowel with simultaneous gastrointestinal stromal tumor

The gastrointestinal tract (GIT) is a common site of metastases for malignant melanoma. These metastatic tumors are often asymptomatic. We describe a case of a 58-year-old male who presented with a sudden onset of generalized abdominal pain. The patient's past medical history was significant for lentigo melanoma of the right cheek. Laparotomy was performed and two segments ofsmall bowel, one with a perforated tumor, the other with a non-perforated tumor, were removed. Histology and immunohistochemical staining revealed the perforated tumor to be a metastatic malignant melanoma and the non-perforated tumor was found to be a gastrointestinal stromal tumor (GIST). The patient was discharged 7 d postoperatively. To the best of our knowledge, this is the first reported case in the literature of a simultaneous metastatic malignant melanoma and a GIST. Surgical intervention is warranted in patients with symptomatic GIT metastases to improve the quality of life or in those patients with surgical emergencies.

Expression of the Tumor Metastatic Suppressor Gene in Mouse Melanoma Model: Inverse Association to Metastatic Potential

Data obtained in experimental cutaneous melanomas have suggested that the nm23 gene may function as a metastasis suppressor gene. The nm23 level in 8 human cutaneous melanoma cell lines and 2 murine melanoma cell lines were examined. Each melanoma cell line was transplanted subcutaneously into the flank of nude mice, and the metastatic behavior was evaluated by counting lung tumor fool and by determining host survival time. It was found that expression of 'm23 mRNA in human melanomas is correlated closely with reduced metastatic behavior in experimental animals and may serve as a sensitive prognostic indicator of malignancy and survival in patients with melanomas.

Antitumor Effect of Cationic INKKI Peptide from Bovine <i>β</i>-Casein on Melanoma B16F10

Cationic peptide with the sequence INKKI 41-45 was isolated from bovine β-casein after tryptic hydrolysis and synthetized. The aim of this work was to evaluate the antiproliferative activity in vitro and antitumor effect in animal model. The in vitro cytotoxicity was evaluated on B16F10 melanoma cells by MTT assay. Detection of apoptosis was measured using the annexin V/PI double staining and cell cycle analysis performed flow cytometry. Caspase-3 activity was analyzed with substrate specific fluorogenic DEVD-MCA. In vivo, antitumor activity was evaluated in B16F10 melanoma tumor-bearing C57BL/6J mice. The animals were treated with 55 mg/kg INKKI administered into peritumoral region, while control group received saline solution. The following antitumor parameters were examined: tumor volume, number of metastases, tumor delayed time, tumor doubling time. Histological analyses were performed with H & E staining. The results showed that INKKI induced dose-response cytotoxicity selective for B16F10 melanoma cells (IC50 1.7 μM) and did not present cytotoxic effects for FN1 fibroblast cells. INKKI-induced apoptosis detected trough of annexin V/PI assay and it was accompanied with an increase of sub-G1 apoptotic fractions and significant increase of caspase-3 cleavage. The tumor-bearing mice treated with INKKI showed a significant reduction in tumor volume of 72.62% and decreased of metastasis number loci. In addition, INKKI caused a significant delay in tumor growth and prolonged the tumor doubling time. Histological analysis revealed an increased of necrosis areas and reduction of tumor cells in tumor treated with INKKI, it was a many hallmark of its antitumor effects observed from in vivo experiments. In conclusion, we show that INKKI is a peptide that could be considered a new putative candidate development to anticancer therapy drug.

Growth of Circulating Tumor Cell-Derived Colonies from Peripheral Blood of Melanoma Patients: Preliminary Characterization of Colony Composition

Circulating tumor cells (CTC) are rarely detected in the blood of cancer patients, even though they are a direct harbinger of eventual patient demise. We developed an innovative CTC culture technology to allow more sensitive isolation, expansion, and characterization of viable colonies from patient blood. In this assay, the entire leukocyte fraction from 10 ml of anticoagulated patient blood is placed into culture medium without any pre-selection. After 16 days in culture, CTC derived colonies are counted. As a proof-of-principle, blood samples from 58 Stage IIa-IV melanoma patients were tested. Ninety percent of these samples grew colonies. The colony numbers ranged from 0 - 308 (mean 63 ± 9.5 SEM). Ten normal volunteers had virtually no growth (mean 0.5 ± 1.4 colonies). Colonies were harvested using a micropipette for characterization. Tumor-cell containing spheroids were embedded in paraffin, sectioned, and stained with melanoma-specific mAb for histologic characterization. MITF proved to be the most consistent immunostain that identified melanoma cells in these colonies. A host-cell component in colonies was also identified using CD68 and CD43 mAb staining. Following enzymatic dissociation of colonies, a variety of immunostains were tested. Papanicolau staining proved most useful for identifying the abnormal nuclei of tumor cells. Flow cytometry could readily distinguish host and tumor cell populations based on DNA content and forward/side scatter in dissociated colonies. The stem cell marker ALDH1A1 associated with the aneuploid population, but CD45 was expressed on both diploid and aneuploid cells. The ability to repeatedly isolate CTC derived colonies from cancer patient blood samples opens the door to a novel type of long-term clinical monitoring. This novel CTC culture technology may prove useful to perform molecular characterization, assessment of treatment response, and testing of drug sensitivity and resistance in patients during treatment.

Melanoma Immunotherapy: Overcoming Obstacles to Augment Anti-Tumor Immune Responses

Melanoma is the most aggressive form of skin cancer and accounts for the vast majority of skin cancer-related deaths. Its ability to metastasize quickly, often before diagnosis, makes this cancer difficult to treat with traditional therapies. The identification of anti-melanoma immune responses in patients and the discovery of tumor antigens targeted by these immune responses have paved the way for immunotherapy as a novel approach to treating this cancer. In this review, the major immunotherapies targeting these melanoma tumor antigens are discussed. The advantages and limitations of peptide-, protein-, and gene-based vaccination maneuvers and adoptive cell transfer therapies are emphasized. Recent insights into melanoma immune evasion strategies are also highlighted, with particular focus on how our increasing knowledge of tumor/immune cell interactions is driving the development of novel immunotherapeutic strategies for the treatment of melanoma.

Myometrium and Sigmoid Colon Metastatic Melanoma Simulating a Ewing Tumor: A Case Report

Malignant melanomas or lymphoma of the skin are malignant tumors of the skin and/or the mucous membranes whose uterine metastases are rare. The secondary uterine localizations, although rare, must be evoked in front of a pelvic tumoral syndrome, or diffuse metastases and a personal past history of melanoma, even after a long time of remission. In our observation, the evolution of the tumor in the pelvis extended to the muscular structures of the uterus and the sigmoid colon of a 72 year old patient, what made it an exceptional case. The diagnosis of these secondary localizations is a diagnosis of elimination, almost always post-operative, made on the histopathological and immunocytochemical study of the surgical specimen, supported by cytogenetics, even molecular biology. The treatment is based on chemotherapy.

Effects of heat shock protein-antigen peptide complexes (HACs) in melanoma B16 cell line on transplanted tumor development in mice

目的 :建立黑色素瘤 B16细胞热休克蛋白 -抗原肽复合物 (HACs)的制备方法并测其抑瘤效应。方法 :应用 Sephacryl S- 2 0 0凝胶过滤制备 HAC粗提物 ,应用 SDS- PAGE纯化 HACs,并测其抑瘤效应。结果 :应用 SDS- PAGE纯化的 HAC6 0、 HAC75和 HAC97不同程度地降低肿瘤发生率 ,延迟肿瘤发生时间和减慢肿瘤生长速度。结论 :6 0 0 0 0～ 970 0 0

MAGEA12在胃肠道间质瘤中的表达及其与危险度分级的关系

目的:探讨胃肠道间质瘤(gastrointestinal stromal tumors,GIST)中黑色素瘤相关抗原基因A12(melanomaassociated antigen gene A12,MAGEA12)的表达水平及其与GIST危险度分级的关系。方法:收集5例GIST患者的肿瘤组织及癌旁组织,采用转录组测序的方法,通过主成分分析(principal components analysis,PCA)、基因集富集分析(gene set enrichment analysis,GSEA)及差异表达基因分析法,明确GIST组织异常信号通路及异常表达基因;采用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)法验证90对GIST组织中异常表达基因MAGEA12的表达水平,并通过χ2检验分析MAGEA12的表达水平与GIST患者临床病理特征的相关性。结果:PCA显示GIST组织基因表达谱与癌旁组织比较差异有统计学意义;GSEA发现,GIST组织中多种肿瘤相关信号通路异常高表达;差异表达基因分析发现,MAGEA12是GIST组织中表达上调最高的基因(表达升高25倍);qRT-PCR验证了MAGEA12在GIST样本中显著高表达,且与GIST危险度分级呈正相关。结论:MAGEA12在GIST中高表达,其表达水平与GIST危险度相关,可能成为治疗的新靶点。

自噬与恶性黑色素瘤侵袭转移相关性研究进展

自噬是真核细胞特有的一种生命现象,对于促进细胞的生存,积极适应不同环境有十分重要的作用。自噬在肿瘤发生和转移的过程中起着双刃剑的作用。在黑素细胞肿瘤中关于自噬的研究结果大不相同,其原因可能与黑素细胞肿瘤的内在异质性以及使用不同的自噬检测技术有关,不同角度的研究均支持自噬与恶性黑素瘤之间存在着密切的相互作用,特别是自噬在药物靶向治疗中的作用更为重要。本文拟综述自噬在黑色素瘤侵袭、转移等生物学过程中的双重作用,为开发恶性黑色素瘤新的治疗方案及发现新的生物标志物提供线索。

PRKCH的表达对CD8^(+)T细胞功能及黑色素瘤免疫治疗的影响

目的 探讨PRKCH表达对CD8^(+)T细胞功能的影响及其在黑色素瘤免疫治疗效果预测中的作用。方法 运用Kaplan-Meier法分析黑色素瘤免疫治疗队列以及肿瘤基因组图谱(TCGA)数据中PRKCH在黑色素瘤中的表达及其与总生存期(overall survival, OS)的相关性。利用单细胞RNA测序(single-cell RNA-sequencing, scRNA-seq)数据分析PRKCH的细胞表达群体,利用蛋白质互作网络(protein-protein interaction networks, PPI)和基因集富集分析(gene set enrichment analysis, GSEA)评估PRKCH诱导抗肿瘤免疫应答的机制。利用逆转录病毒过表达和敲减PRKCH,检测小鼠CD8^(+)T细胞功能。利用B16-OVA小鼠黑色素瘤模型验证PRKCH过表达的OT-1 CD8^(+)T细胞对肿瘤生长的影响。结果 在黑色素瘤免疫治疗队列和TCGA数据中,高水平的PRKCH显著延长患者OS(P<0.05)。scRNA-seq分析显示PRKCH可以在T细胞表达。PPI、GSEA和scRNA-seq分析显示PRKCH的高表达与更多的细胞毒性和记忆性T细胞形成相关。过表达和敲减PRKCH分别增强和减少CD8^(+)T细胞的增殖及IFN-γ、Granzyme B的表达。过表达PRKCH增强OT-1 CD8^(+)T细胞的肿瘤抑制能力。结论 PRKCH增强CD8^(+)T细胞抑制肿瘤进展的能力,并可以用于预测黑色素瘤免疫治疗效果。

黑色素瘤循环肿瘤细胞检测的金属卤化物钙钛矿外泌体复合探针检测新策略

液体活检技术的兴起为黑色素瘤的快速、准确诊断提供了新的机遇。然而,普通循环肿瘤细胞活检基于上皮黏附蛋白进行阳性富集,但信号标记的有机荧光探针存在量子效率低的问题,导致检测黑色素瘤循环肿瘤细胞时准确率和灵敏度较低。本文以高量子效率的金属卤化物钙钛矿量子点作为信号标记物,以黑色素瘤来源的外泌体作为生物识别分子,构建了一种用于黑色素瘤液体活检的循环肿瘤细胞检测新策略。与商品化的上皮细胞黏附蛋白富集策略相比,本研究报道的复合探针检测新策略,其检测灵敏度提高了一个数量级,并且具有良好的亲水性和低毒性。实验结果证明了外泌体引导的金属卤化物钙钛矿量子点指示的黑色素瘤循环肿瘤细胞检测新策略具有理想的应用前景。

铁死亡相关基因作为皮肤黑色素瘤预后因素的多组学分析

目的:黑色素瘤具有高度恶性和异质性。开发特定的黑色素瘤预后预测模型对提高患者的生存率和选择治疗策略至关重要。最近,铁死亡已被证明是一种由过度脂质过氧化诱导的铁依赖性程序性细胞死亡形式。然而,铁死亡相关基因(ferroptosis-related genes,FRGs)与黑色素瘤预后的相关性仍不清晰。本研究评估FRGs在黑色素瘤中的作用,开发一种新的预后模型,旨在为黑色素瘤的个性化治疗及疗效改善提供新思路。方法:首先通过系统地表征癌症基因组图谱(The Cancer Genome Atlas,TCGA)-皮肤黑色素瘤(skin cutaneous melanoma,SKCM)中73个FRGs的遗传改变和mRNA表达。同时通过反转录聚合酶链反应和蛋白质印迹法验证筛选的特定靶基因。随后使用TCGASKCM队列构建多基因特征模型。根据特征模型将黑色素瘤患者分为高风险和低风险组,对铁死亡相关的特征模型与免疫特征、免疫治疗的疗效或药物反应进行相关分析。结果:通过分析TCGA-SKCM数据集中的黑色素瘤样本,发现FRGs在基因变异和拷贝数变异方面表现出高频率,这些变化显著影响了基因的表达。此外,与正常皮肤组织相比,在黑色素瘤组织中发现了30个显著差异表达的基因。随后使用LASSO Cox回归方法构建的FRGs相关预后模型成功识别了13个与患者总体生存预后相关的FRGs,并通过外部数据集验证了该模型的有效性。最后,功能富集和免疫响应结果分析进一步表明高风险和低风险组之间存在免疫细胞浸润、突变负担和低氧状态的显著差异,且该模型能有效预测免疫治疗响应和药物敏感性。结论:本研究建立了一种强预后预测模型,可为黑色素瘤患者的靶向治疗和免疫治疗提供新的方向。

单细胞转录组测序在黑色素瘤中的研究进展

黑色素瘤是皮肤癌中致死性最高的一种恶性肿瘤,其发生、发展机制复杂,具有高度侵袭性、转移性和致死性,患者预后差。因此,加强该病基础研究尤为重要。单细胞转录组测序作为近年来快速发展的新技术,以其特有的优势如探索细胞亚群的异质性、鉴定稀有细胞、使用生物信息算法揭示不同组织细胞的分化轨迹等,极大地推动了黑色素瘤的基础与临床研究发展。本文就黑色素瘤单细胞转录组测序研究中肿瘤异质性和肿瘤免疫方面的进展进行综述。

嵌合型抗原受体T细胞在恶性黑色素瘤中应用进展

恶性黑素瘤细胞膜表达有多种生物标志物,可作为治疗靶点开发针对性的嵌合型抗原受体T细胞(CAR-T),CAR-T目前对恶性黑素瘤中GD2、HER2、CD70、Gp100、NYESO-1等靶点已表现出良好的疗效。本文旨在梳理恶性黑素瘤CAR-T靶点及临床进展,分析工程化T细胞的挑战与应对之策,以期CAR-T应用于恶性黑素瘤临床实践。

肿瘤标志物及EMT通路表达水平对黑色素瘤预后的影响

目的探讨上皮-间充质转化(EMT)通路对黑色素瘤患者预后的影响及黑色素瘤常见肿瘤标志物与EMT通路的相关性,为临床识别和随访高转移风险的黑色素瘤患者提供新思路。方法从TCGA数据库中获得461例黑色素瘤患者的转录及临床数据,通过GSVA包对每个样本的表达进行非参数的无监督分析,得到EMT通路的表达评分并分为低和高两组,使用Survival包进行生存分析。随后对肿瘤标志物的mRNA和EMT通路表达水平进行Spearman相关性分析,并根据年龄、性别和肿瘤分期进行亚组分析。进一步从TCGA数据库中下载黑色素瘤患者肿瘤标志物的蛋白表达水平,同样进行Spearman相关性分析,从mRNA和蛋白表达水平2个角度探讨常见肿瘤标志物与EMT通路的相关性。最后将上述肿瘤标志物、EMT通路表达水平和临床特征纳入多因素分析以明确对黑色素瘤预后的影响。结果EMT通路的高表达预示黑色素瘤患者的不良预后。黑色素瘤肿瘤标志物PMEL、MLANA、TYR和MITF的mRNA表达水平与EMT通路呈负相关(P<0.05),MKI67的mRNA表达水平与EMT通路呈正相关(P<0.05)。肿瘤标志物与EMT通路相关性的亚组分析结果与总样本中所得结果基本一致。肿瘤标志物PMEL高表达和EMT相关通路低表达时预示良好预后,EMT相关通路低表达时MLANA、MKI67低表达预示良好预后(P<0.05)。PMEL、MLANA和MITF的蛋白表达水平与EMT通路的表达呈负相关(P<0.05)。PMEL、年龄和病理分期是影响黑色素瘤预后的独立危险因素(P<0.05)。结论黑色素瘤的常见肿瘤标志物PMEL、MLANA和MITF较低表达可能与EMT通路的高表达相关;EMT相关通路低时,PMEL与MLANA高表达、MKI67低表达预示良好预后,随访中应给予一定程度重视。

直肠肛管区特殊类型肿瘤的诊治进展

直肠肛管区存在诸如直肠神经内分泌肿瘤、胃肠间质瘤、肛管鳞癌、直肠肛管黑色素瘤、原发性直肠淋巴瘤等特殊类型肿瘤,其临床特征与直肠癌存在差异,且发病率较低,导致临床医师对其认识不足。本文对直肠肛管区特殊类型肿瘤的临床诊疗进展进行综述。

Improving recognition of hepatic perivascular epithelioid cell tumor:Case report and literature review

A 58-year-old man presented with the chief complaint of abdominal bloating and was incidentally found to have a liver tumor.As diagnostic imaging studies could not rule out malignancy,the patient underwent partial resection of segment 3 of the liver.The lesion pathologically showed eosinophilic proliferation,in addition to immunohistochemical positivity for human melanoma black 45 and Melan-A,thereby leading to the diagnosis of a hepatic perivascular epithelioid cell tumor(PEComa).A PEComa arising from the liver is relatively rare.Moreover,the name ‘PEComa' has not yet been widely recognized,and the same disease entity has been called epithelioid angiomyolipoma(EAML),further diminishing the recognition of PEComa.In addition,PEComa imaging findings mimic those of malignant liver tumors,and clinically,this tumor tends to enlarge.Therefore,a PEComa is difficult to diagnose.We conducted a systematic review of PEComa and EAML cases and discuss the results,including findings useful for differentiating perivascular epithelioid cell tumors from malignant liver tumors.

Comparative Study on the Inhibitory Effect of RecombinantFN Polypeptide CH50 and CH56 on the Metastasis ofMelanoma Cells

On the basis of preparation of anti-metastatic recombinant FN polypeptides, CH50 and CH56, we further studied the function of these polypeptides.The capacity of CH50 binding with melanoma cells (ED50 30 mM) was higher than that of CH56 (ED50 45 mM). Both of the polypeptides could significantly suppress the binding of melanoma B16 cells to laminin. There was no significant difference in the inhibitory effect between two polypeptides. In the experimental metastasis of melanoma cells, both of CH50 and CH56 could significantly inhibit the metastasis of the tumor cells, and reduce the number of lung metastasis by about 80%. Our results suggest that Ⅲ-11 and ED-A repeats influenced, to some extent, the binding capacity of bifunctional-domain polypeptide to cells, but did not affect the inhibition of the polypeptide on the metastasis of melanoma cells. The presence and connection of cell Ⅰ and Hep Ⅱ domains are the elements which determine the ability of recoinbinant FN polypeptides to inhibit the metastasis of tumor cells.

Primary gastric melanoma: A case report with imaging findings and 5-year follow-up

BACKGROUND Most melanomas identified in the stomach are metastatic;primary gastric melanoma(PGM)is extremely rare,and the relevant studies are relatively scarce.PGM may be incorrectly diagnosed as other gastric malignant tumor types.CASE SUMMARY We describe a rare case of PGM confirmed through long-term clinical observation and pathological diagnosis.A 67-year-old woman presented to our hospital with recurrent chest tightness and chest pain.Digital gastrointestinal radiography revealed a circular shadow in the gastric cardia.Computed tomography(CT)revealed a heterogeneous tumor with uneven enhancement.Enlarged lymph nodes were noted in the lesser curvature of the stomach.On magnetic resonance imaging(MRI),T1-and T2-weighted imaging revealed hyperintensity in and hypointensity in the tumor,respectively,both of which increased substantially after uneven enhancement.Near total gastrectomy was performed,and the tumor was pathologically confirmed to be a gastric melanoma.Because no other possible primary site of malignant melanoma was suspected,a clinical diagnosis of PGM was made.The patient was followed for nearly 5 years,during which she received CT reexamination,but no recurrence or metastasis was observed.CONCLUSION Certain imaging characteristics could be revealed in PGM.Imaging examination can be of great value in preoperative diagnosis,differential diagnosis,and followup of patients with PGM.

Novel anti-melanoma treatment:focus on immunotherapy

Melanoma is an intractable cancer that is aggressive, lethal, and metastatic. The prognosis of advanced melanoma is very poor because it is insensitive to chemotherapy and radiotherapy. The incidence of melanoma has been ascending stably for years worldwide, accompanied by increasing mortality. New approaches to managing this deadly disease are much anticipated to enhance the cure rate and to extend clinical benefits to patients with metastatic melanoma. Due to its high degree of immunogenicity, melanoma could be a good target for immunotherapy, which has been developed for decades and has achieved certain progress. This article provides an overview of immunotherapy for melanoma.